Process of manufacturing a vaccin for the prevention of hog-cholera and product therefrom.



in a ear CHARLES WARREN DUVAL, OF NEW ORLEANS, LOUISIANA, ASSIGNOR 0F TWO-THIRJDS T0 MAURICE J'. COURET, OF NEW ORLEANS, LOUISIANA.

PROCESS OF MANUP'ACTURING A VACCIN FOR THE PREVENTION OF IIOG-CHOLEBA AND PRODUCT THEREFROM No Drawing.

Therefrom; and I do hereby declare the following to be a full, clear, and exact descrlption of the invention, such as will enable others skilled in the art to which it appertains to make and use the same.

My invention relates to the manufacture of a vaccin for the prevention of hog cholera, and it consists in a process whereby a'vaccin is secured which, when properly applied, is safe andreliable to protect the hog for a period of at least a year, and probably indefinitely, against hog cholera.

My invention also includes the product of said process, as will be hereinafter more fully described.

I take from a hog which is in the advanced state of hog cholera, and therefore heavily infected with the virus, the blood, fiesh, brain, heart, liver, spleen and kidneys and collect them under strict aseptic con-v removing, as far as possible, theclitions, ligaments, capsules, vessels, etc. These organs and flesh are then passed through an ordinary meat grinder under aseptic conditions, and the mass thus obtained is thoroughly mixed with the blood taken from the infected hog. This mixture is then frozen by or through the agency of carbon dioxid snow, or other equivalent method, so as to facilitate its further,pulverization. This is accomplished by trituratingthat is to say, by mashing at l grinding in a mortar, or some similar container. This trituration is done while the mass is in thefrozen state. The mixture is then placed in glass or other suitable containers, and is then rapidly desiccated in vacuo at approximately freezing temperature (0 C.) over some neutral hygroscopic agent, such as sulfuric acid, caustic soda, or unslaked lime. During the process of desiccation, the water is completely removed, and there is left a residue of solid material in a finely divided condition. This residue is then ground to a fine powder under aseptic conditions. This powder contains the living virus of hog "This'produces in Specification of Letters Patent. Patented 1113, 26,191

Application filed March 22, 1916. Serial N0. 85,960.

cholera, whose infectivity has not been appreciably altered ment. The viability of the virus of' hog cholera is thus preserved for such use as may be desired, and the possibility of its alteration or destruction through contaminating germs is eliminated. This powder is then transferred to air-tight containers and stored in'a refrigerator maintained at or about freezing temperature (0 C.)', under which conditionkits infectivity is maintained and found to be preserved unaltered for the entire period-a year or more during which the product has been under observation.

The powderproduced in the above manner will hereinafter to as the virus powder and it will be mixed, as .will be hereinafter described, with a second powder which will be hereinafter called the antitoxict powder, the mixture being the vaccin product.for which I now claim a patent. The antitoxic powder is produced as follows: A normal hog is taken and there is injected into this hog, in proper increasing virus of hog cholera, either in the form of the virus powder, above referred to, or in the fresh state (the virus powder being cheaper and more economically produced and preserved than the virus in the fresh state).

such a hog a high immunity. This immunity should be raised by the method aforesaid 'to that point at least where five milligrams of inactivated and subsequently desiccatedserum from this animal will sensitize one milligram of the virus powder above described. Proper standardization must be had as in all bacteriological products. The blood of the hog, thus immunized, is drawn from the hog fromtime to time, as required, due regard being had always to draw the blood at a time when the immunity is at proper height? If the immunity be not at proper height, the immunity should be heightened as and to the extent hereinabove set forth. The blood so drawn from the hog is first defibrinated, and then it is centrifugalized, or permitted to stand for a sufficiently long time-say 24 hours. By this process of centrifugalization, or standing, the heavy material in the blood settles, and the clear serum remains on the top; it is then drawn off into other conbe called and referredv while, and until necessary forpractical use,

tainers. The clear serum thus obtained is inactivated at a temperature of approxidesiccated rapidly in vacuo at approximately freezing temperature (0 C.) over some neuv .tral hygroscopic agent such as sulfuric acld,

caustic soda or unslaked lime. The potency of the desiccated powder thus obtained can be maintained for a' year or more, and possibly indefinitely, if kept at approximately freezing temperature (0 (1.). The virus powder maintained at approximately freezing temperature (0 C.) is then mixed with the antitoxic powder likewise maintained at ap roximately freezing temperature (0 C 'in the following proportions and manner, to wit The proportion of this mixture is one part by weight of virus powder to approximately fifty parts by weight of antitoxic powder. To obtain a more uniform mixture of these powders, the virus powder is emulsified in sterile waterin the proportions of one quarter of one cubic centimeter (0.25 C. C.) of water to one milligram (1 mg.) of virus powder. Add to this emulsion the proper proportion of antitoxic powder, as above set forth, and again thoroughly emulsify. This mixture is then sub jected to a temperature of approximately 37 C. for an hour, or to room temperature (20-25 C.) for from six to twelve hours. By this latter method, sensitization of'the virus powder is effected, This mixture, or sensitized emulsion, is then desiccated rapidly in vacuo at approximately freezing temperature (0 C.) over some neutral hygroscopic agent, such as sulfuric acid, caustic soda or unslaked lime. The powder thus obtained is the finished product, or vaccin, and it is ready for administration. Meanit is kept in sealed glass containers at approximatelyifreezing temperature (0 0.). When needed for practical administration,

the product is. mixed with some suitable vehicle such as glycerin, and applied to' a pointed instrument for the purpose of vaccination; or it may be suspended in sterile water and injected in the hog hypodermatically; or it may be injented into .the hog hypodermatically in the form of apellet. This product is not infectious in any quantity. The dose thereof for protection against the natural disease (hog cholera) is 50 milligrams; in-localities wherethe disease exists in more virulent form, larger doses may be used with advantage. By increasing the dose and repeating the administration at suitable intervals, the immunity of the hog can be heightened or raised and brought t that point known as hyperimmunization,

when its blood can be drawn off and usedin the production of commercial antitoxin.

In the. hereinbefore described method the tenuation is 1 with antitoxin powder? but similar results may be obtained by attenuating or killing mately 56 C. for thirty minutes, and is then the virus in the virus inafter described.

The proportion by weight of virus pow- ,der and antitoxic powder may be changed from that of one to fifty, as hereinabove powder, as will behereset forth, by previouslyattenuating or de-- straying the viability of the virus'powder, and then mixing the attenuated or killed virus powder with the antitoxic powder, following the method above described. This attenuation or destruction of viability is accomplished by subjecting the virus powder, suspended in sterile water, to a temperature of from 10 C. to 60 C. for periods ranging, respectively, from four months to one hour. By these processes of either attenuation or killing, theinfectivity of the the virus powder, so suspended, be subjectedto a, temperature of approximately 38 C. for a period of four weeks, such ati ewise accomplished. Again, if the virus powder, so suspended, be subjected to a temperature of approximately "60 C. for a period of approximately one hgur its viability is destroyed. Within these limits the attenuation of the virus powder, or the destruction of its viability, may be accomplished by proper experimentation,

and the relative temperature and time fixed.

When such attenuated or killed virus powdertis used in mixture with the antitoxic powder, following the hereinabove described method of sensitization, the proportion by weight of attenuated or killed virus powder to the antitoximpowder varies from one to forty to-one to five, de-' v pending uponthe degree of attenuation up to the complete destruction of viability The proper-proportions within these limits must be determined by. standardization.

The dose is the same whether the product be made with the virulent virus powder or with'the attenuated or killed virus powder; and .the'administration of the vaccine is had in the same way.

The -hereinbefore described methods provide for-the previous desiccation of the virus mixing the virus powder, either virulent,

attenuated or ln'lled, with the antito'xic,

powder, it is equally practicableto mixfthe and the antitoxic serum before bringing. "them mtoadmixture. However, instead of portion of onemilli am of virus owder to one-half of 1 cublc'centimeter .C.) of

- said serum. materials are then thoroughly mixed, and sensitization effected by subjecting to the temperature, as in the manner hereinabove set forth, after which the mixture is rapidly desiccated in vacuo at approximately freezing temperature (0. C.) over some neutral hygroscopic agent, such 'as sulfuric acid, caustic soda, or unslaked lime.

It will be noted that the sensitized virus may be employed in its liquid condition without being finally reduced to the desiccated or powdered stateas above described; but I prefer to reduce the product to the final desiccated state for convenience in handling and administering same.

This completed dried product, like the completed product obtained by the mixture of the virus powder with the antitoxic powder is returned to approximately freezing temperature (0 C.) until necessary for practical administration, and it may be administered in the same dose and manner as hereinabove described. Thus, it will be seen that I use the virus powder, either in the virulent, attenuated, or killed state, with either the antitoxic serum in the powdered state, or in the fresh, or liquid, state.

I claimas my invention.

1. The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog parts containing the virulent virus, desiccating the mass so secured, and converting same into the powdered form, and finally mixing this product with inactivated antitoxic serum, and sensitizing the same.

2. The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog parts containing the virulent virus, desiccating the mass so secured, and converting same into the owdered form, attenuating the virus of the d s iccated powder so secured, and finally'lmlxing this product with inactivatedantitoxic serum, and sensitizing the same.

3. The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog parts virus, desiccating the mass so secured, and converting same into the powdered form, killing the virus of the desiccated powder so secured, and finally mixing this product with inactivated antitoxic serum, and sensitizing the same.

4. The process for the production of a vaccin product for the prevention of hog 63 cholera which consists in removing from the infected hog parts containing the virulent virus, converting same into the powdered form, and finally mixing this product while sus- 65 pended in sterile water with inactivated antider so secured,

containing the virulent desiccating the mass so secured, and

toxic serum, also in the desiccated and pow" dered form, and sensitizing the same.

5. The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog parts containing the virulent virus, desiccating themass so secured, and

converting same into the powdered form, attenuating the virus of the desiccated pow and finally mixing this product, while suspended in sterile water, with inactivated antitoxic serum, also in the desiccated and powdered form, and sensitizing the same.

6. The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog parts containing the virulent virus, desiccating the mass" so secured, and converting same into the powdered form, killing the virus of the desiccated powder so secured, and finally mixing this product, while suspended in sterile water, with inactivated antitoxic serum, also in the desiccated and powdered, form, and sensitizing the same.

7. The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog parts containing the virulent 5 virus, desiccating the mass so secured at a temperature of approximately 0 0., and converting same while at substantially the same temperature into the powdered form, mixing the product with inactivated antio toxic serum, and then sensitizing the same.

8. The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog parts containing the virulent virus, desiccating the mass so secured at a temperature of approximately 0 (3., and converting same while atasubstantially the same temperature into the powdered form, attenuating the virus of the desiccated pow- 11o der so secured, mixing the product with in activated antitoxic serum, and then sensitizing the same.

9. The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog parts containing the virulent virus, desiccating the mass so secured at a temperature of approximately 0 CI, and converting same while at substantially the 12o same temperature into the powdered form, killing the virus of the. desiccated powder s o secured, mixing the product with inactlvated antitoxic serum, and then sensitizing the same. a

10. Theprocess for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog parts containing the virulent virus, desiccating the mass so secured at a 130 fit vaccin product for tion of the-virus is produced, and finally desiccating the sensitized mass.

11. The process for the production of vaccin product for the prevention of hog cholera, which consists in removing from the I infected hog parts containing the virulent virus, desiccating the mass so secured at a -t'emperature of approximately 0 0., and converting same while at substantially the same temperature into the powdered form, then securing from thenblood' of a highly immunized hog the clear serum, inactivating same by subjecting itto a temperature of '.'.5 approximately 56 C. for substantially minutes, desiccating this serum and reducing themass to the powdered form, then suspending the two powders in suitable proportions. in sterile water, subjecting the emul- "30 sion to a temperature of approximately 37 C. for approximately one hour, whereby sensitization of the" virus is produced, and finally desiccating the sensitized mass.

12. The process for the production of a vaccin product for the prevention of hog cholera whichc nsists in removing-from the {infected hog parts containiiig the virulent I virus, desiccating the mass so secured ata temperature of approximately 0 (3., con- 40 verting same while at substantially the same temperature into t e powdered form, and attenuating the virus of the desiccated powder so secured; then securing from the blood of perature of approximately 56 C. for substantially 30 minutes, desiccating this serum and reducing the mass/to the powdered form; then suspending the two powders in suitable proportions in sterile water, sub'- g ject'ing the emulsion to a temperature of approximately 37 C. for approximately one hour, whereby sensitization of the virus prbduced; and finally desiccating the sensitized mass. I x 14. The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the infected hog arts containing the virulentvirus, desiccatlng the mass so secured at a temperature of approximately 0 0., converting same while at substantially the same temperature into the powdered form, and

' killing the virus of the desiccated powder so secured; then securing from the blood of a highly immunized hog the clear serum, inactivating same by subjecting it to a temperature of lapproximately 56 G or sub stantially 30 minutes; then mixing this latter product with the virus powder, subject- 'ing the mixture to a temperature of approximately 37 C. for approximately one hour, whereby sensitization of thevirus is produced; and finally desiccating the sensitized mass. v J

15. The process for the productionof a vaccin product for the prevention of hog.

' cholera which consists in removing from the "a highly immunized hog the clear serum, inv 15 activating same by subjecting it-to a temperature of approximately'56 C.- for substantially 30 minutes; then mixing: this latter product with the virus -powder, 'subjectingthe mixture to a temperature of approxi- 50-mately 37 C. for approximately one'hour,

whereby sensitization of the virus is pro- 4 duced, and finally desiccating the sensitized 7. mass;

13. The. process for the production of a the prevention of hog infected" hog parts containing the virulent virus, desiccating the mass so secured at'a m0 temperature of approximately 0 (1, converting same while at substantially the sametemperature into the powdered'form, and killing the Virus of the desiccated powder so securedythen securing from the blood ofa highly immunized hog the clear serum, inactivating same by subjecting it to atem perature' of approximately 56 C. for sub-K stantially 30 minutes, desicc'ating this serum and ,reducing the mass to the powdered form; then suspending the two powders in suitable proportions in :sterile water, sub- I jecting the emulsion to a temperature of ap-" vcholera which consists in removing from the infected hog parts, containing the virulent virus, desiccating the mass so, secured ata temperatureof approximately 0 "0., con-' 60 Q verting same 'Whileat substantially the same temperature into the powdered fprm, and'at- I tenuating the virus of the desiccated powder, I

so securedythen securing from the bloodof a highly immunized hog the clear serum,.i1 1- .mixing'thisproduct with inactivated anti proximately 37 C. for approximately one hour, whereby sensitization of the virus is 1 5 produced; and finally desiccating the sensitized mass.' g

16. The process for the production of a vaccin productfor the prevention of hog cholera which consists in removing from the 1 20 infected, hog parts containing the virulent virus, desiccating the mass so secured, and convertmgsame mto the powdered form,

toxic-serum, sensitizing'thesame, and finally ldesiccating the mass and reducing the same 65 activating sameby. subjecting it to 'aYtemto the powdered form. I

' 17., The processnfor' the production of a vaccin product for 'theprevention of hog cholera which cohsists iii fr emoving from the uct with inactivated antitoxic serum, sensivaccin pro tizing the same, and finally desiccating the mass and reducingthe same, to the powdered form.

18. The vaccin product for the prevention of cholera which consists in removing from the infected hog parts containing the virulent process for the production of) a virus, desiccating the mass so secured, and

converting same into the powdered form, killing the virus of the desiccated powder so secured, and finally --mixing this product ,with inactivated antitoxic serum, sensitizing the same, and finallydesiccating the mass. and reduclngthe same to the powdered then mixing this product, while suspended in sterile water, with inactivated antitoxic serum, also in the desiccated and powdered form, sensitizing the same, and finally desiceating the mass and reducing the same to the powdered form.

20 The process for the production of a vaccin product for the prevention of hog cholera which consists in removing from the 1 infected hog parts containing he virulent virus, desiccating the massso secured, converting same into the powdered form, attenuating the virus of the desiccated powder so secured, then mixing this product, while suspended in sterile water, with inactivated antitoxic-serum, also in the desiccated and powdered form, sensitizingithe. same, and finally desiccating the mass and reducing the same'to the powdered form.

21. The

not "for the prevention of hog cholera which consists in removing from the infected hog parts containing the virulent virus, desiccating the mass so secured, converting sameinto the powdered form, killing the virus of the desiccated powder so secured, then mixing this product, while suspended in sterile water, with inactivated antitofic serum, also in the desiccated and blood or other soft tissues of by mixing the same with process for the production of 5- powdered form, sensitizing the same, and

finally desiccating the mass and reducing the same to the owdered form.

22. A pr not for the prevention of hog cholera comprising a vaccin material made from the desiccated product of any one or more of the blood or other soft tissues of a hog infected with hog cholera, subsequently sensitized by mixing thesame with the inactivated antitoxic serum of an immunized 0g. v

23. 'A product for the prevention of hog cholera comprising a vaccin material made fromthe desiccated product of any one or more of the blood or other soft tissues of a hog infected with hog cholera, subsequently attenuated and thereafter sensitized by mixing the same with the inactivated antitoxic serum of an immunized hog.

24. A product for the prevention of hog cholera comprising ,a vaccin material made from the desiccated product of any one or more of the blood or othersoft tissues of a hog infected with hog cholera, the viability of which virus has been destroyed, and thereafter sensitized by mixing the same with the inactivated antitoxic serum of an immunized hog. I

25. A product for the prevention of hog cholera comprising a vaccln material in the form of a dry powder, made from the desiccated product of any one or more of the a hog infected with hog cholera and subsequently sensitized the inactivated antitoxic serum of an immunized hog.

cholera comprislng a vaccin material in the form of a dry powder, made-from the desiccated product of any one or more of the blood or other soft tissues of a hog infected with hog cholera and subsequently attenuated ,and thereafter sensitized by mixing the same with the inactivated antitoxic serum of an immunized hog.

27. A product for the prevention of hog cholera comprising a vaccin material in the form of a dry powder, made from the desiccated product of any one or more of the blood or other soft tissues of a hog infected with hog cholera, the viability of which virus has been destroyed and thereafter sensitized by mixing the same with the inactivated antitoxic serum of an immunized hog.

In testimony whereof I aflix my signature.

CHARLES WARREN DUVAL. 

